Event Title

Session 3E: Detecting AB, ABO, APP, p-Tau, and t-Tau in Chick Retina

Session Number

Session 3E: 1st Presentation

Advisor(s)

William Klein and Kirsten Viola, Northwestern University

Location

Room A113

Start Date

28-4-2017 1:15 PM

End Date

28-4-2017 2:30 PM

Abstract

The purpose of this investigation was to detect AB (Amyloid beta), ABOs (Amyloid Beta Oligomers), APP (Amyloid Precursor Protein), p-Tau, and t-Tau in the chick embryo retina. This would determine if the chick embryo was suitable for Alzheimer’s disease research. In this investigation, chick eggs were incubated for 12-13 days, after which they were dissected and the brain and retina were removed. These were then homogenized and dotted triplicate onto nitrocellulose membrane strips. A dot blot was performed on these strips, and the results were imaged using a Kodak imaging station. This process was repeated multiple times with slight variations each time in order to result in the most optimal images. The AB, ABOs, APP, p-Tau, and t-Tau were detected after performing a dot blot. This occurred after determining that the lack of protein and excess of secondary antibody was causing a lack of detection. To remedy this, a dot blot concentrator was used to increase protein levels, and the proper dilutions of secondary antibody, 1.0 ug/mL NU2 and 1:40000 parts anti-rabbit, were determined.

Comments

Additional team members: Kristen Schuster

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Apr 28th, 1:15 PM Apr 28th, 2:30 PM

Session 3E: Detecting AB, ABO, APP, p-Tau, and t-Tau in Chick Retina

Room A113

The purpose of this investigation was to detect AB (Amyloid beta), ABOs (Amyloid Beta Oligomers), APP (Amyloid Precursor Protein), p-Tau, and t-Tau in the chick embryo retina. This would determine if the chick embryo was suitable for Alzheimer’s disease research. In this investigation, chick eggs were incubated for 12-13 days, after which they were dissected and the brain and retina were removed. These were then homogenized and dotted triplicate onto nitrocellulose membrane strips. A dot blot was performed on these strips, and the results were imaged using a Kodak imaging station. This process was repeated multiple times with slight variations each time in order to result in the most optimal images. The AB, ABOs, APP, p-Tau, and t-Tau were detected after performing a dot blot. This occurred after determining that the lack of protein and excess of secondary antibody was causing a lack of detection. To remedy this, a dot blot concentrator was used to increase protein levels, and the proper dilutions of secondary antibody, 1.0 ug/mL NU2 and 1:40000 parts anti-rabbit, were determined.